IL29925A - Tetanus vaccine and process for preparing it - Google Patents

Description

AND PROCESS FOB PREPARING IT mas Tetanus vaccine and process for preparing it 5265 A Ma 86 The present invention relates to a tetanus vaccine and to a process for preparing The tetanus vaccines contain inactivated tetanus the tetanus Vaccines of this kind sometimes cause vaccination and No method has been described hitherto to eliminate these The present invention provides a tetanus vaccine which has been obtained by treating tetanus obtained in known manner by cultivation of tetanus bacteria and subsequent lation and inactivation of the with pepsin in the sence of at a in the range of from to preferably to 5 purifying it in known manner by means of a protein precipitating agent and working it up to As starting material for the production of the vaccine of the present there may be used tetanus toxoid which has been obtained in known manner by cultivation of tetanus isolation and inactivation of the toxin is contained in the known vaccines Ullmanns Encyclopadle technischen Volume page 611 the tolerance of these toxoids is proved by the process of the present Since the tanus toxoid is not soluble in the indicated acid the treatment with pepsin according to the present invention is carried out in the presence urea in an amount for example about preferably 5 mols of In urea the tetanus toxoid is also soluble the for example fluence on the ferment activity of the The treatment is suitably carried out with a pepsin concentration of to preferably 1 of mg of tetanus The duration of the action of the pepsin on toxoid is mined by a preliminary It on the aggregation gree of the toxoid and is in the range of from 6 to 60 preferably 24 to 48 After the pepsin which is carried out at a temperature in the range of from 0 to preferably at body temperature the active material is precipitated by means of a known protein precipitating preferably the same volume of a saturated ammonium sulfate in order to separate first the low molecular weight gradation after dissolution the it is fractionated in two the first precipitate obtained up to an ammonium sulfate saturation of 22 is inactive and is therefore whereas the second which is obtained up to a saturation of 50 contains the treated tetanus For the there may also be used other known protein precipitating for example ethanol or Before being used as a the tetanus toxoid modified according to the present invention must still be filtered under sterile It may be used as it may be injected after combination with an for example aluminium hydroxide or aluminium Compared to the known tetanus the vaccine of the present invention has a considerably better it is highly active storable for at least one year at C without loss of The tolerance of the vaccine prepared according to the present invention is a ed son and the metrically determined diameter of the local nation Another proof of the tolerance of the vaccine was obtained by active anaphylactic tests on When Guinea pigs are with an antigen and the same antigen is stered intravenously to them after two or three they suffer a shock and This is likewise the case when the tanus toxoid of the state of the art or a tetanus vaccine which is available in the commerce is used as the All of ten Guinea pigs treated in such manner the Guinea pigs immunized with tetanus toxoid were injected vaccine of the present a considerably better rance was out of ten Guinea six survived the described anaphylaxis The activity of the of the present invention was determined by reinforcing When in an acute for example in the case of it is necessary to rapidly obtain a high antibody a reinforcing vaccination or a booster vaccination is applied By booster there is to be understood the increase of immunity with steep of the titer by a reinforcing vaccination after a basic immunization effected some time test was effected on l l test In this a tetanus toxoid obtained in known manner by cultivation of tetanus bacteria and following isolation and inactivation of the toxin by means of was compared with the vaccine of the present All test persons had already been immunized against The tetanus toxoid contained 10 the vaccine of the Lf ml b Lf Limes there is to be understood the quantity of antigen which flocks out 1 of specific All test persons ceived a dose of 5 The following table shows the tive units of of serum of the test after reinforcing vaccination with the tetanus toxoid and the vaccine of the present titer classes and is thus considerably better than that of the tetanus it is to be noted that this rior activity is already obtained by ml with only half the quantity of antigen which had been contained in tetanus toxoid solution The vaccine of the present invention is administered muscularly or deep The vaccine of the present invention shows sedimentation coefficient ranging between about S to 100 and S to 0 The following Examples illustrate the invention but they Example 100 of a tetanus toxoid obtained by cultivation of tetanus bacteria in known manner and subsequent isolation and inactivation of the the protein content of which amounted to and the of which had been adjusted to by means of about ml of acetic were combined with g of urea and then 15 mg of pepsin and the solution was incubated at The duration of the action was determined by a preliminary It depends on the degree of aggregation of the toxoid and was in the present case 24 hours solution was then adjusted to pH by means of about of combined with the same about ml of a saturated ammonium sulfate the toxoid it was isolated by dissolved in 50 of a physiological NaCl solution and again combined with about ml of a saturated ammonium sulfate This precipitate was The decanted portion was dialyzed until free from salt and Preliminary test Samples were taken after hours and tested for their degree of degradation by Since the size of the molecules is decisive for the speed of migration in the starch gel owing to the sieving the electrophoresis can be used for proving the reduction of the size the In this it was found the vaccine of the present invention migrated faster than the starting Aggregations which were contained in the starting which manifested in electrophoresis by a slower migrating The reduction of the size of the molecules could also be proved by measurement of the sedimentation coefficient with the aid of an it was found that the starting material had a sedimentation coefficient of whereas the product of the present invention had a sedimentation coefficient of Example 2 100 ml of tetanus whose protein content amounted to and whose was adjusted by means of about ml of binormal acetic were dialyzed against molar acetate buffer having a of and bound to urea in a molar ratio of 1 and containing of sodium 15 mg of pepsin were added and the solution was cubated for 28 hours at The degradation product was then purified in a manner analogous to that described in Example insufficientOCRQuality

Claims (1)

1. CLAIMED A teta containing tetanus toxoid sedimentation coefficients of between 5 3 S to and S to 0 that has been treated with pepsin in the presence of urea in a a of 1 5 to A tetanus as claimed Claim 1 containing a tetanus toxoid that has been treated with to 0 rag of mg of tetanus toxoid in a urea solution containing of urea per A process fo preparing a tetanus vaccine as claimed i Claim 1 wherein a tetanus in known by cultivation of tetanus bacteria and following isolation and of the is treated with pepsin in the presence of urea in a of from 5 to purified in know manner by means of a protein precipitating and worked u to a A process as claimed Claim urea concentration of 3 6 per liter is A process claimed Claims 3 or wherein 5 to mg of of insufficientOCRQuality